简易微流控芯片技术在生理流动条件下体外动态分析阿司匹林和氯吡格雷对健康志愿者血小板黏附聚集的影响  

Dynamical Analysis of the Inhibitory Effect of Aspirin and Clopidogrel on Platelet Adhesion and Aggregation for Healthy People under Physiological Flow Condition by Microfluidic Chip Technology

作　　者：陈静[1] 丁玲 何翠[1] 陈丹 邓素容[1] 龚放[1] 李远[1] CHEN Jing;DING Ling;HE Cui;CHEN Dan;DENG Surong;GONG Fang;LI Yuan(Central Laboratory,Yongchuan Hospital,Chongqing Medical University,Chongqing 402160,China;Yongchuan Sub-center,Chongqing Blood Center,Chongqing 402160,China)

机构地区：[1]重庆医科大学附属永川医院中心实验室,重庆402160 [2]重庆市血液中心永川分中心,重庆402160

出　　处：《中国医学科学院学报》2018年第3期299-307,共9页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(11702047);重庆市社会事业与民生保障科技创新专项(cstc2017shms A130009);重庆市卫生和计划生育委员会医学科研项目(2017MSXM079);重庆医科大学附属永川医院院级重点项目(YJZQN201534)~~

摘　　要：目的采用简易微流控芯片技术在体外动态分析生理流动条件下阿司匹林和氯吡格雷对健康志愿者血小板黏附聚集的影响。方法随机招募12名健康志愿者,采集其外周静脉血液样品并分别用20μmol/L乙酰水杨酸(ASA)、50μmol/L 5-磷酸-2-甲基硫代腺苷酸(2-Me SAMP)、20μmol/L ASA+50μmol/L 2-Me SAMP处理,以未处理血液样品作为对照组。将血液样品以1000 s^(-1)动脉生理性相关剪切率流过Ⅰ型胶原蛋白修饰微通道300 s,同时采用显微镜动态拍摄荧光标记血小板在胶原蛋白表面黏附聚集的荧光图像,以血小板覆盖率作为血小板黏附聚集行为的量化指标。结果在1000 s-1剪切率流动条件下,对照组血样在胶原蛋白表面可观察到符合体内预期的血小板黏附聚集动态行为;阿司匹林或氯吡格雷单独处理抑制流动后期(200~300 s)血小板黏附聚集行为,而阿司匹林和氯吡格雷联合处理则降低流动中前期(≤150 s)血小板黏附数量并降低流动后期(200~300 s)血小板聚集体稳定性;阿司匹林和氯吡格雷联合处理对血小板黏附聚集行为抑制具有协同效应;12名志愿者在对阿司匹林和氯吡格雷抑制血小板聚集响应上表现出异质性。结论简易微流控芯片技术可为抗血小板药物效应分析提供一种新的技术平台。Objective To explore the inhibitory effect of aspirin and clopidogrel on platelet adhesion and aggregation behaviors under the physiological flow condition using microfluidic chip technology for healthvolunteers. Methods Peripheral venous blood samples collected from twelve randomly recruited health volunteers were treated with 20 μmol/L acetylsalicylic acid,50 μmol/L 2-methlthioadenosine-5＇-monophosphate triethylammonium salt,and their combination,respectively,with untreated blood samples being control group. The blood samples were flowed through a microchannel modified with type I collagen protein at a physiological relevant shear rate of 1000 s-1 for 300 s,while the fluorescent images of platelet aggregations were dynamic captured using a microscope. Based on the images,the platelet coverage rates were calculated and used as quantitative parameters for evaluating platelet adhesion and aggregation behaviors. Results Under a flow condition of 1000 s^（-1） shear rate,an expected in vivo-like platelet adhesion and aggregation behaviors were observed at the surfaces of collagen proteins for control blood samples. Aspirin alone or clopidogrel alone suppressed platelet adhesion and aggregation at the later period of flow（200-300 s）,while the combination of aspirin and clopidogrel reduced the adhesion numbers of platelets at the earlier stage of flow（ ≤150 s） and compromised the stability of platelet aggregation at the later period of flow（200-300 s）. The combination showed synergistic effect in inhibiting platelet aggregation. Furthermore,such inhibitory effect was heterogeneous among 12 volunteers. Conclusion This simple microfluidic technology can offer a new technical platform for analyzing the inhibitory effect of antiplatelet drugs.

关 键 词：血小板聚集 阿司匹林 氯吡格雷 微流控芯片 流动剪切率 

分 类 号：R-331[医药卫生]